Silkworm recombinant bovine zona pellucida protein 4 (bZP4) as a potential female immunocontraceptive antigen; impaired sperm-oocyte interaction and ovarian dysfunction

Porcine zona pellucida proteins (ZPs) have been utilized as female immunocontraceptive antigens. The purpose of this study was to explore the potential use of silkworm recombinant bovine ZP4 as an alternative. When the protein was injected with monophosphoryl lipid A (MPL) – an immuno-stimulative agent – into two female goats, marked elevation of the anti-ZP4 titer was detected. Application of the purified specific IgG to a porcine in vitro fertilization system reduced the sperm penetration rate. In one goat, the cyclic profile of serum progesterone disappeared as the anti-ZP4 titer increased. Histological examination of the ovaries revealed degeneration of antral follicles with sparse infiltration of inflammatory cells in the theca, indicating that autoimmune oophoritis had been induced. Together, the present results suggest that recombinant ZP4 disturbs fertilization and exerts a pathogenic effect on follicle development in goats, thus indicating its potential as a female immunocontraceptive antigen.     

Abnormal clonalities of B-lymphocytes in bovine leukemia virus-infected cattle with persistent lymphocytosis

Peripheral B-lymphocyte clonality of 274 bovine leukemia virus-infected cattle with lymphocytosis was analyzed using clonality PCR based on sequences of the variable region of the bovine immunoglobulin H chain. None of the cattle showed monoclonal proliferation, while 10, 31, and 233 showed minor-clonal, oligoclonal, and polyclonal proliferation, respectively. A total of 163 cattle were analyzable the following year, and lymphocytosis was maintained in 157, indicating persistent lymphocytosis (PL). B-lymphocyte clonality of the 157 PL cattle was minor-clonal in 6 (3.8%), oligoclonal in 8 (5.1%), and polyclonal in 143 (91.1%). A higher rate of enzootic bovine leukosis (EBL) onset within a year was observed in PL cattle with minor-clonal (50.0% (3/6)) and oligoclonal (25.0% (2/8)) proliferation compared to those with polyclonal (5.6% (8/143)) proliferation. Minor-clonal and oligoclonal proliferation in PL cattle may be a prognosis factor for developing EBL.     

Comparison of Physical Properties of Wheat Starch Gels with Different Amylose Content

The effects of amylose content and other starch properties on concentrated starch gel properties were evaluated using 10 wheat cultivars with different amylose content. Starches were isolated from grains of two waxy and eight nonwaxy wheat lines. The amylose content of waxy wheat lines was 1.4–1.7% and that of nonwaxy lines was 18.5–28.6%. Starch gels were prepared from a concentrated starch suspension (30 and 40%). Gelatinized starch was cooled and stored at 5°C for 1, 8, 16, 24, and 48 hr. The rheological properties of starch gels were studied by measuring dynamic viscoelasticity with parallel plate geometry. The low‐amylose starch showed a significantly lower storage shear modulus (G′) than starches with higher amylose content during storage. Waxy starch gel had a higher frequency dependence of G′ and properties clearly different from nonwaxy starches. In 40% starch gels, the starch with lower amylose showed a faster increase in G′ during 48 hr of storage, and waxy starch showed an extremely steep increase in G′. The amylose content and concentration of starch suspension markedly affected starch gel properties.     

Mixed Thymoma in a Young Cynomolgus Monkey (Macaca fascicularis)

A mass with a diameter of 1.5 cm was found in the thymus of a 4-year and 3-month-old male cynomolgus monkey. Microscopically, the mass consisted of two different patterns of proliferation, dense or fascicular proliferation of elongated spindle cells in a sporadic storiform pattern and dense proliferation of thymic cortex-like lymphoid cells in which the multifocal pale nests resembling the thymic medulla were distributed. In these pale nests, large dendriform cells sometimes forming Hassall's corpuscles were present. The proliferating spindle cells were positive for cytokeratin. The lymphoid cells in the mass were positive for CD3. We concluded that the mass consisted of the neoplastic thymic epithelium with thymocytes proliferation containing medullary differentiation. The mass was diagnosed as a mixed thymoma according to the WHO classification of thymomas in humans. Mixed thymoma is characterized as a mixture of two types of proliferative lesions, spindle-shaped epithelial proliferation and a lymphocyte predominant lesion with or without medullary differentiation. To the best of our knowledge, this is the first report concerning thymoma in monkeys.     

Assessing the potential impacts of climate change on the alpine habitat suitability of Japanese stone pine (Pinus pumila)

To assess the potential distribution of Pinus pumila, a dominant species of the Japanese alpine zone, and areas of its habitats vulnerable to global warming, we predicted potential habitats under the current climate and two climate change scenarios (RCM20 and MIROC) for 2081–2100 using the classification tree (CT) model. The presence/absence records of P. pumila were extracted from the Phytosociological Relevé Database as response variables, and five climatic variables (warmth index, WI; minimum temperature for the coldest month, TMC; summer precipitation, PRS; maximum snow water equivalent, MSW; winter rainfall, WR) were used as predictor variables. Prediction accuracy of the CT evaluated by ROC analysis showed an AUC value of 0.97, being categorized as “excellent”. We designated Third Mesh cells with an occurrence probability of 0.01 or greater as potential habitats and further divided them into suitable and marginal habitats based on the optimum threshold probability value (0.06) in ROC analysis. Deviance weighted scores revealed that WI was the largest contributing factor followed by MSW. Changes in habitat types from the current climate to the two scenarios were depicted within an observed distribution (Hayashi’s distribution data). The area of suitable habitats under the current climate decreased to 25.0% and to 14.7% under the RCM20 and MIROC scenarios, respectively. Suitable habitats were predicted to remain on high mountains of two unconnected regions, central Honshu and Hokkaido, while they were predicted to vanish in Tohoku and southwestern Hokkaido. Thus P. pumila populations in these regions are vulnerable to climate change.     

Epidemiological study of Kabuto Mountain virus,a novel uukuvirus,in Japan

Kabuto Mountain virus (KAMV), the new member of the genus Uukuvirus, was isolated from the tick Haemaphysalis flava in 2018 in Japan. To date, there is no information on KAMV infection in human and animals. Therefore, serological surveillance of the infection among humans and wild mammals was conducted by virus-neutralization (VN) test and indirect immunofluorescence assay (IFA). Sera of 24 humans, 59 monkeys, 171 wild boars, 233 Sika deer, 7 bears, and 27 nutria in Yamaguchi Prefecture were analyzed by VN test. The positive ratio of humans, monkeys, wild boars, and Sika deer were 20.8%, 3.4%, 33.9% and 4.7%, respectively. No positive samples were detected in bears and nutria. The correlation coefficients between VN test and IFA in human, monkey, wild boar, and Sika deer sera were 0.5745, 0.7198, 0.9967 and 0.9525, respectively. In addition, KAMV was detected in one pool of Haemaphysalis formosensis ticks in Wakayama Prefecture. These results indicated that KAMV or KAMV-like virus is circulating among many wildlife and ticks, and that this virus incidentally infects humans.     

Primary epididymis squamous cell carcinoma in a CB6F1-Tg rasH2 mouse

In a 26-week carcinogenicity study in rasH2-Tg mice, squamous cell carcinoma on the epididymis was observed in a male mouse in the positive control group treated with N-methyl-N-nitrosourea. A 29-week-old male rasH2-Tg mouse that was euthanized 21 weeks after the administration of N-methyl-N-nitrosourea had a white-grayish mass on the left caput epididymis. The mass was nodular and consisted of pleomorphic tumor cells forming alveolar, sheeted, and trabecular structures suggesting epithelial tumor growth. These cells presented a cobblestone-like arrangement and formed intercellular bridges. Keratinization was infrequently observed. Periodic acid-methenamine-silver staining revealed argyrophilic fibrous structures around the alveolar structure of the tumor cells. Immunohistochemically, the tumor cells were positive for cytokeratin AE1/AE3 and cytokeratin 14 and negative for cytokeratin 5, p63, uroplakin III, vimentin, desmin, and αSMA. These immunohistochemical results suggested the tumor cells originated from the epididymal ducts. Metastatic lesions were observed in the mesenteric, inguinal, and pancreaticoduodenal lymph nodes. Based on these results, this tumor was diagnosed to be a primary squamous cell carcinoma of the epididymis. This is the first report of primary squamous cell carcinoma of the epididymis in a rasH2-Tg mouse.     

Analysis of Potassium Uptake by Rice Roots Treated with Aluminum Using a Positron Emitting Nuclide, 38K

We investigate the effect of Al on K⁺ uptake by rice roots. Potassium-38 (³⁸K), a positron emitting nuclide (the half-life: 7.61 min), was used to trace K⁺ behavior. When a rice root was treated with 10μM Al for 24 h, the uptake of ³⁸K in the root was increased in the range of 1 to 2 cm from the root tip compared with that of the control sample. Because the root continued to grow without showing any damage of plasma membrane during the Al treatment, it was suggested that the ³⁸K uptake was not occurred through diffusion into the cells. The uptake of ³⁸K in both treatments, with/without Al, was decreased by VO₄^3- (inhibitor of H⁺-ATPase on plasma membrane) and DNP (H⁺ ionophore) treatment, which suggested that the K⁺ uptake was performed through an active transport, such as H⁺:K⁺ transport or H⁺ gradient promoted by an Al treatment.     

Effect of added organic matter on soil fertility after stripping-off Cs-contaminated top soil at Iitate village in Fukushima Prefecture

The agricultural fields were contaminated by the radionuclides ¹³⁴Cs and ¹³⁷Cs after the nuclear power plant accident in Fukushima. Prior to the accident, local farmers had successfully established sustainable agriculture in Iitate Village using natural farming practices and recycling. Since 2011, decontamination work such as stripping-off the top soil has been ongoing on agricultural land. Although decontamination is essential, it could cause an unfortunate decrease in soil fertility. Here, we examined the use of organic matter as a means to quickly recover the fertility of the agricultural top soil. We transplanted rice crops into three paddy plots: one received rice straw that had been harvested there last year, another received composted manure, and the third (control) received no additives after decontamination. We applied 40 kg/10a of basal fertilizer and 20 kg/10a of KCl each plot. The rates of Cs concentration in unhulled rice/rice straw were around 0.001. Tendency of plant heights increase and leaf chlorophyll content decrease were similar in the three treatment plots. However, the numbers of stems on 111 days after the transplant were 21, 15, and 19, unhulled rice yield were 513, 462, and 310 g/m², in the rice straw, cattle manure compost, and control plots, respectively. Soil properties of three plots were similar. Radioactive Cs concentrations in the new rice from each treatment plots were lower than the maximum allowed level set by the Japanese government. These results revealed that treating soil with rice straw might have great potential to aid the recovery of a paddy field after stripping-off the top soil. Notably, this treatment significantly improved the yield of rice and supplied organic matter without additional labor.     

Integrating agroecology and landscape multifunctionality in Vermont: An evolving framework to evaluate the design of agroecosystems

Agroecosystems cover vast areas of land worldwide and are known to have a large impact on the environment, yet these highly modified landscapes are rarely considered as candidates for landscape design. While intentionally-designed agricultural landscapes could serve many different functions, few resources exist for evaluating the design of these complex landscapes, particularly at the scale of the whole-farm. The objective of this paper is to introduce an evolving framework for evaluating the design of agroecosystems based on a critical review of the literature on landscape multifunctionality and agroecology. We consider how agroecosystems might be designed to incorporate additional functions while adhering to agroecology principles for managing the landscape. The framework includes an assessment tool for evaluating farm design based on the extent of fine-scale land use features and their specific functions, to consider the present state of the farm, to plan for future conditions, or to compare alternative futures for the design of the farm. We apply this framework to two farms in Vermont that are recognized locally as successful, multifunctional landscapes. The Intervale Center, an agricultural landscape located within the city limits, serves as an incubator for new farm startups and provides unique cultural functions that benefit the local community. Butterworks Farm, a private operation producing organic yogurt and other food products, achieves important ecological functions through an integrated crop-livestock system. These farms and many others in Vermont serve as models of a framework that integrates landscape multifunctionality and agroecology in the design of the landscape. In the discussion section, we draw from the literature and our work to propose a set of important themes that might be considered for future research.